Hypothalamo-pituitary disconnected (HPD) and control Soay rams were treated chronically for 48 wk with s.c., continuous-release implants of melatonin while under long days (16L:8D). The implants produced continuously elevated blood concentrations of melatonin 2-3 times higher than the normal nocturnal maximum. The long-term treatment induced a biphasic effect on prolactin secretion in both the HPD and control rams, with a marked decrease in the blood prolactin concentrations for 10 wk followed by a gradual increase. The introduction of a second melatonin implant after 20 wk failed to affect prolactin secretion. The treatment with melatonin also caused a dynamic effect on FSH secretion, but this occurred in the control rams only. Blood concentrations of FSH in the HPD rams were very low throughout, but there were minor changes in testicular diameter that were correlated with variations in prolactin. Overall, the results support the conclusion that 1) melatonin acts primarily in the pituitary gland to affect prolactin secretion, and partial refractoriness develops at this level for control of prolactin; and 2) melatonin acts most probably in the hypothalamus to affect gonadotropin secretion, and refractoriness develops at the level of the neural tissue regulating GnRH release for control of gonadotropins.